Overview | ![]() PrinterFriendlyVersion |
Ex/Em(nm) | 540/590 |
MW | N/A |
CAS# | N/A |
Solvent | N/A |
Storage | F/D/L |
Category | NeuroBIOLOGy ReactiveOxygenSpecies |
Related | CellSignaling CellFunctionalAnalysis BiochemicalAssays |
1.Preparecells:
1.1 Foradherentcells:Platecellsovernightingrowthmediumat10,000to40,000cells/well/90μLfora96-wellplateor2,500to10,000cells/well/20μLfora384-wellplate.
1.2 Fornon-adherentcells:CentrifugethecellsfromtheculturemediumandsUSPendthecellpelletsinculturemediumat100,000-200,000cells/well/90µLfora96-wellpoly-Dlysineplateor25,000-50,000cells/well/20µLfora384-wellpoly-Dlysineplate.Centrifugetheplateat800rpmfor2minuteswithbrakeoffpriortoyourexperiment.
Note:Eachcelllineshouldbeevaluatedonanindividualbasistodeterminetheoptimalcelldensity.
2.PrepareMitoROS™580workingsolution:
2.1 PrepareMitoROS™580stocksolution(500X):Add50µLofDMSO(ComponentC)intothevialofMitoROS™580(ComponentA),andmixthemwell.
Note:25μLofreconstitutedMitoROS™580stocksolutionisenoughfor1plate.Unusedportioncanbealiquotedandstoredat<-20°Cformorethanonemonthifthetubesaresealedtightlyandkeptfromlight.Avoidrepeatedfreeze-thawcycles.
2.2 PrepareMitoROS™580workingsolution:Add25μLof500XDMSOreconstitutedMitoROS™580stocksolution(fromStep2.1)into10mLofAssayBuffer(ComponentB),andmixthemwell.Thisworkingsolutionisstableforatleast2hoursatroomtemperature.
3.Runsuperoxideassay:
3.1 Treatcellswith10μLof10Xtestcompounds(96-wellplate)or5μLof5Xtestcompounds(384-wellplate)inyourdesiredbuffer(suchasPBSorHHBS).Forcontrolwells(untreatedcells),addthecorrespondingamountofcompoundbuffer.
3.2 Toinducesuperoxide,incubatethecellplateat37°Cforadesiredperiodoftime,protectedfromlight.
Note:WetreatedHeLacellswith50µMAntimycinA(AMA)at37ºCfor30minutestoinducesuperoxide.SeeFigure1fordetails.
3.3 Add100μL/well(96-wellplate)or25μL/well(384-wellplate)ofMitoROS™580workingsolution(fromStep2.2)intothecellplate.
3.4 Incubatethecellsat37°Cfor30minto1hour,andmonitorthefluorescenceincreaseatEx/Em=540/590nm(cutoff=570m)withbottomreadmode,ortakeimagesusingfluorescencemicroscopewithaTRITCfilter.
References&Citations | ![]() PrinterFriendlyVersion |
1. KitamuraK,MaruyamaK,HamanoS,KishiT,KawakamiT,TakahashiY,OnoderaS.(2014)Effectofhypochloriteoxidationoncholinesterase-inhibitionassayofacetonitrileextractsfromfruitsandvegetablesformonitoringtracesoforganophosphatepesticides.JToxicolSci,39,71.
2. MartinDE,DeAlmeidaJF,HenryMA,KhaingZZ,SchmidtCE,TeixeiraFB,DiogenesA.(2014)Concentration-dependenteffectofsodiumhypochloriteonstemcellsofapicalpapillasurvivalanddifferentiation.JEndod,40,51.
3. YinB,DengJ,PengX,LongQ,ZhaoJ,LuQ,ChenQ,LiH,TangH,ZhangY,YaoS.(2013)Greensynthesisofcarbondotswithdown-andup-conversionfluorescentpropertiesforsensitivedetectionofhypochloritewithadual-readoutassay.Analyst,138,6551.
4. TakimotoK,TaharaguchiM,SakaiK,TakagiH,TohyaY,YamadaYK.(2013)Effectofhypochlorite-baseddisinfectantsoninactivationofmurinenorovirusandattempttoeliminateorpreventinfectioninmicebyadditiontodrinkingwater.ExpAnim,62,237.
5. SetlowB,YuJ,LiYQ,SetlowP.(2013)AnalysisofthegerminationkineticsofindividualBacillussubtilissporestreatedwithhydrogenperoxideorsodiumhypochlorite.LettApplMicrobiol,57,259.
6. ThornRM,RobinsonGM,ReynoldsDM.(2013)Comparativeantimicrobialactivitiesofaerosolizedsodiumhypochlorite,chlorinedioxide,andelectRochemicallyactivatedsolutionsevaluatedusinganovelstandardizedassay.AntimicrobAgentsChemother,57,2216.
7. Perez-CruzF,CortesC,AtalaE,BohleP,ValenzuelaF,Olea-AzarC,SpeiskyH,AspeeA,LissiE,Lopez-AlarconC,BridiR.(2013)Useofpyrogallolredandpyranineasprobestoevaluateantioxidantcapacitiestowardshypochlorite.Molecules,18,1638.
8. ZhangJ,YangX.(2013)Asimpleyeteffectivechromogenicreagentfortherapidestimationofbromateandhypochloriteindrinkingwater.Analyst,138,434.
9. CollaoB,MoralesEH,GilF,PolancoR,CalderonIL,SaavedraCP.(2012)DifferentialexpressionofthetranscriptionfactorsMarA,Rob,andSoxSofSalmonellaTyphimuriuminresponsetosodiumhypochlorite:down-regulationofrobbyMarAandSoxS.ArchMicrobiol,194,933.
10. ZhangJ,WangX,YangX.(2012)ColorimetricdeterminationofhypochloritewithunmodifiedgoldnanoparticlesthroughtheoxidationofastABIlizerthiolcompound.Analyst,137,2806.
AAT Bioquest常用产品报价单
产品名 | 货号 | 公司分类 | 商城分类 | 美金价 |
AAT Bioquest/Fluorescein, disodium salt *CAS 518-47-8*/2/100 mg | 2 | iFluor Rapid Tests | 酸碱缓冲液 | 1088.00 | AAT Bioquest/14-3-3 ζ (Ab-58) Antibody/8B0001/50 ug | 8B0001 | Signaling Intermediate Ab | 重组抗体 | 2828.00 | AAT Bioquest/Opioid Receptor (Phospho-Ser375) Antibody/8A0022/50 ug | 8A0022 | Phospho-Specific Ab | 受体 | 2828.00 | AAT Bioquest/iFluor™ 350 goat anti-mouse IgG (H+L)/16440/200 ug | 16440 | Fluorescent Anti-IgGs | 荧光染料 | 1088.00 | AAT Bioquest/trFluor™ Eu goat anti-mouse IgG (H+L)/16518/100 ug | 16518 | Fluorescent Anti-IgGs | 抗小鼠 | 4278.00 | AAT Bioquest/RPE-streptavidin conjugate/16900/100 ug | 16900 | Streptavidin Conjugates | 其他生物染料 | 1378.00 | AAT Bioquest/GCNT3 Antibody/8C14708/50 ug | 8C14708 | Signaling Intermediate Ab | 功能性抗体 | 2828.00 | AAT Bioquest/Amplite™ Luciferase Reporter Gene Assay Kit *Bright Glow*/12518/1 plate | 12518 | Reporter Gene Enzymes | 其它检测试剂盒 | 2103.00 | AAT Bioquest/ReadiLink™ Rapid mFluor™ Violet 450 Antibody Labeling Kit *Microscale Optimized for Labeling 50 &mi | 1100 | mFluor™ Dyes and Kits | 标记试剂盒 | 2103.00 | AAT Bioquest/Amplite™ Colorimetric Urea Quantitation Kit *Blue Color*/10058/200 Tests | 10058 | Diagnostic Molecules | 定量试剂盒 | 2828.00 | AAT Bioquest/Screen Quest™ CHO-Gqi Chimera Cell line/38101/Each | 38101 | cAMP GPCR Assays | 常规细胞株 | 0.00 |